Sperm Preparation for Intrauterine Insemination Using Density Gradient Separation
|
|
- Olivia Sherman
- 5 years ago
- Views:
Transcription
1 Sperm Preparation for Intrauterine Insemination Using Density Gradient Separation 14 Ashok Agarwal, Sajal Gupta, and Rakesh Sharma 1 Introduction Sperm washing is performed to remove seminal plasma and increase sperm density prior to intrauterine insemination (IUI) [ 1 ]. Gradient sperm washing is highly effective at isolating motile sperm from the ejaculate of most men. IUI may be indicated in cases of poor postcoital testing due to hostile or absent cervical mucus, oligospermia, or the presence of sperm antibodies [ 2 ]. It may be used in conjunction with artificial insemination or IVF procedures. 2 Specimen Collection The patient is instructed on how to collect the specimen (see Semen Collection and Labeling Procedure ). The patient collects the specimen into a sterile container and brings it to the laboratory. The patient, or partner, is told to return in ~1 h to pick up the washed specimen. 3 Equipment and Materials A. Components of the sperm separation kit sterile colloidal suspension of silica particles stabilized with covalently bound hydrophilic silane supplied with HEPES-buffered human tubal fluid (HTF): 1. Lower phase (80 %) 2. Upper phase (40 %) A. Agarwal, PhD, HCLD (ABB), ELD (ACE) (*) S. Gupta, MD (Ob/Gyn), MS (Embryology), TS (ABB) R. Sharma, PhD Andrology Center and Reproductive Tissue Bank, American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH, USA agarwaa@ccf.org; guptas2@ccf.org; sharmar@ccf.org B. Sperm washing media (modified HTF with 5 % human serum albumin and gentamicin C. Disposable sterile 15 ml polystyrene conical centrifuge tube(s) with cap(s) D. Centrifuge E. 37 C incubator F. Sterile graduated serological pipettes (2 ml) G. Computer-assisted semen analyzer (CASA) H. Long stemmed (9 ) pasteure pipette I. Disposable 20 μm sperm counting chamber J. Makler counting chamber K. Dilution cups 2-mL L. Brown paper bags M. Disposable sterile transfer pipettes N. Viscosity treatment system (when applicable)-5 mg chymotrypsin 4 Quality Control A. Sperm recovery and percent motility are checked and recorded semiannually on all IUI samples. Criteria : None defined. B. Daily Precision: A patient specimen should be selected at random and run through CASA. A manual count and motility reading should be performed simultaneously. Criteria : All manual results should be within 20 % of CASA value. Response : If results are not within the defined percentage difference, the sample must be repeated. If it is still out-of-range, inform supervisor or director. C. Technologists review patient results to check for technical and clerical errors prior to release of specimen for insemination. Springer International Publishing Switzerland 2016 A. Agarwal et al. (eds.), Andrological Evaluation of Male Infertility, DOI / _14 101
2 102 5 Procedure 5.1 Prepare Reagents 1. Bring the upper and lower phase, HTF, and semen sample to 37 C by incubating for 20 min (Fig ). 2. Transfer 2 ml of the lower phase into a sterile disposable 15 ml conical centrifuge tube. 3. Layer 2 ml of the upper phase on top of the 2 ml lower phase using a sterile pipette. Slowly dispense the upper phase lifting the pipette up the side of the tube as the level of the upper phase rises. Note : A distinct line separating the two layers should be observed. This two-layer gradient is stable for up to 2 h. A. Agarwal et al. *Note : Color-coded labeling tape should be used for all tubes, media, and paperwork and should be specific (of the same color) for each patient. 4. Have the person delivering the specimen sign the Specimen Drop-off/Pick-up Form. They must at this time present an acceptable ID card (e.g., driver s license, employee badge). The technologist accepting the specimen must record on the sheet the type of identification presented and the appropriate ID number. 5. Label a 2 ml conical cup for the post-wash analysis. Remove the warm tube of sperm wash media (HTF with 5 % HSA) from the 37 C incubator and label it with the patient s name and colored labeling tape* (same color as used above). 5.2 Prepare Paperwork/Accept Patient Specimen 1. Make sure the specimen container is labeled with two identifiers. Acceptable identifiers are either the patient name and date of birth or the patient name and medical record number (or SSN). Note : If all of the patient information on the specimen cup is not present, the container should be labeled in front of the patient. 2. Have the Sperm Wash Worksheet, Artificial Insemination by Husband, Specimen Drop-off/Pick-up Form, and two IUI labels prefilled before the patient arrives with all pertinent information. 3. Label a 15 ml centrifuge tube with the patient s and partner s name, medical record numbers, and date. The tube should also be labeled with color-coded tape* as an extra identifier. 5.3 Wash/Analyze Specimen Note : Always use sterile technique during specimen processing. 1. Allow the semen specimen to liquefy completely for ~20 min in a 37 C incubator before processing. Note : Occasionally, samples do not liquefy properly and remain too viscous to pass through the gradient. Add 5 mg of chymotrypsin to the viscous sample ~5 10 min before layering to increase motile sperm yields. 2. Using a sterile transfer pipette, place a few drops of the patient s semen sample into a conical cup. Place ~5 μl of the sample onto a sperm counting chamber and microscopically verify that there is motile sperm before proceeding with the wash (this sample can also be used for the prewash analysis). Fig Incubator set at 37 C and depiction of sample undergoing liquefaction [Reprinted with permission, Cleveland Clinic Center for Medical Art & Photography All Rights Reserved]
3 14 Sperm Preparation for Intrauterine Insemination Using Density Gradient Separation Measure the total semen volume using a sterile 2 ml serological pipette. 4. Gently layer no more than 3 ml* of liquefied semen onto the upper phase. *Note : If the sample volume is greater than 3 ml, it is necessary to split the specimen into two tubes for processing. Process the second tube using the same procedure that was used for the first tube. Combine the second tube with the first tube during step #11 after adding HTF media. 5. Centrifuge for 20 min at 1600 rpm (Fig ). 6. While the specimen is in the centrifuge, perform a prewash CASA analysis. Perform CASA analysis according to the Routine Semen Analysis protocol and record the results on the worksheet. Note : While examining the specimen, pay particular attention to any extraneous round cells, debris, and bacteria that are present. If round cells are greater than or equal to 1.00 M/mL, perform an Endtz test immediately. A positive Endtz test should be reported to the appropriate medical personnel as soon as possible. The Endtz result should also be written on the tube s outer label at the end of the wash procedure before handing the specimen to the patient. 7. After the specimen has centrifuged, remove the supernatant without disturbing the pellet. Using a sterile long pasteure pipette carefully remove the seminal plasma, upper interphase (raft a ), upper (40%) colloid layer, and the lower interphase raft b ). Leave the majority of the 80% colloid layer in place and discard the aspirated material. 8. Using another clean pasteure pipette remove the soft pellet by direct aspiration (maximum 0.5 ml) from the bottom of the tube beneath the lower (80%) colloid layer into the soft. Avoid contaminating the pasteure tip aperture with the residual seminal plasma/raft material. 9. Transfer the pellet to a clean 15 ml conical tube. 10. Using a serological pipette, add 2 ml of HTF media and resuspend the pellet by pipetting the sample gently up and down until the sperm pellet is properly mixed into the sample. 11. Centrifuge the resuspended sample for 7 min at 1600 rpm (Fig ). 12. Repeat step 7 above (Fig ). 13. Resuspend the final pellet (Fig ) using 0.5 ml of HTF media with a 2 ml serological pipette. Note the volume of the sample. Leave a small droplet (~20 μl) in a labeled conical cup for the post-wash analysis. Subtract this volume from the volume noted above and record on the patient worksheet (Fig ). 14. Seal the tube using tamper-evident tape, label it with the premade label, and place it in the incubator until the patient arrives. Show the color-coded tube to the patient and have the patient verify the couple s names, medical record numbers, and date. Place the sample in a brown paper bag and hand it to the patient. Be sure the patient signs the specimen pick-up form and presents appropriate identification. The patient should carry the specimen to gynecologist for the insemination. The gynecologist determines whether or not the sperm count and motility are sufficient for Fig Double-density gradient wash procedure; separation of seminal plasma, abnormal nonmotile sperm, and viable motile sperm [Reprinted with permission, Cleveland Clinic Center for Medical Art & Photography All Rights Reserved]
4 104 A. Agarwal et al. Fig HTF resuspended sample centrifuged to produce viable sperm pellet [Reprinted with permission, Cleveland Clinic Center for Medical Art & Photography All Rights Reserved] insemination. If no motile sperm are found on the post-wash specimen, page the gynecology nurse prior to releasing specimen. Note : Always look at the post-wash specimen on a sperm counting chamber before releasing it to the patient. Notify the inseminator immediately if bacteria are present. If there is a positive Endtz test, report the result to the nurse as soon as possible and write it on the outside label of the tube before handing it to the patient. If the prewash Endtz is positive and the post-wash round cell count is less than 1 M/mL, note the round cell count on the outside label of the tube. 13. Perform a post-wash CASA analysis according to the Routine Semen Analysis protocol and record the results on the patient worksheet. Page the nurse with the patient name, clinic number, total motile sperm, Endtz value (if necessary), and any other pertinent information. 14. Record appropriate information (below) on the Artificial Insemination by Husband form: (a) MRN (b) Date of insemination (c) Total motile post-wash sperm (d) Wash type (e.g., gradient) (e) Insemination performed (Gyn nurse) (f) Comments (e.g., positive Endtz, agglutination, high viscosity) (g) Tech initials 15. After checking for technical and clerical errors, enter the results and print the final report. Check the final report to ensure the accuracy of the final results. Note : Immediately correct any inaccurate results.
5 14 Sperm Preparation for Intrauterine Insemination Using PureCeption 105 Fig Removal of supernatant HTF [Reprinted with permission, Cleveland Clinic Center for Medical Art & Photography All Rights Reserved] Fig Sperm pellet resuspended in HTF [Reprinted with permission, Cleveland Clinic Center for Medical Art & Photography All Rights Reserved] 6 Procedural Notes A. It is important to use sterile plasticware and glassware during the sperm wash procedure. B. Each patient specimen should be kept in a separate specimen rack during the time of processing. C. Processing of patient specimen(s) should be started and completed by a single technologist. In rare cases, when a second technologist is called to help during the processing, the patient s paperwork should be reverified by the assisting technologist. 7 Tips on How to Maximize the Yield from Density Gradient A. Make sure you have all components of the gradient brought to 37 C temperature before use. Besides avoiding a cold shock to the spermatozoa, condensation on the media bottles disappears allowing for better inspection of the bottles for contamination. Any bottle whose contents appear in any way cloudy or hazy should not be used. B. Do not use the same pipette in more than one bottle of media. C. Density gradient components are air-buffered. Prolonged exposure to a 5 % CO 2 environment will alter the ph of these products, which may in turn affect their nature and performance. D. For highly viscous semen add 5 mg of chymotrypsin to the ejaculate 10 min before it is placed on top of the gradient increases motile sperm yields without any measurable damage to the motile sperm. E. Do not overload the gradient. The volume of semen in this case is less important than the absolute number of sperm placed on the gradient. Usually, a maximum number should be around 120 million cells. Overloading the gradient will result in a phenomenon called rafting this aggregation of desirable, as well as undesirable, components of the semen will be
6 106 created and will be present in the post-centrifugation pellet. F. Always use the gradient within 1 h after creating it eventually the two phases over time blend into each other and a sharp interface will not exist. G. To optimize the number of motile sperm, avoid contamination with the seminal plasma raft a or raft b, or the upper and lower layers. H. Use a long stem preferably a 9 Pasteure pipette to carefully aspirate the soft pellet at the bottom of the tube beneath the 80% layer. I. Transfer the final soft pellet into a fresh 15 ml conical tube for second step of centrifugation. References A. Agarwal et al. 1. Beydola T, Sharma RK, Lee W, Agarwal A. Sperm preparation and selection techniques. In: Rizk B, Aziz N, Agarwal A, Sabanegh E, editors. Male infertility practice. New Delhi: Jaypee Brothers Medical Publishers; Agarwal A, Allamaneni S. Artificial insemination. Section 6: infertility and recurrent pregnancy loss. Clinical reproductive medicine and surgery: Tommaso Falcone and William Hurd. Elsevier publishers.
7 14 Sperm Preparation for Intrauterine Insemination Using Density Gradient Separation 107
Cryopreservation of Client Depositor Semen
Cryopreservation of Client Depositor Semen 16 Ashok Agarwal, Sajal Gupta, and Rakesh Sharma 1 Introduction The primary purpose of the Andrology Center and Reproductive Tissue Bank is to provide quality
More informationHUMAN TUBAL FLUID MEDIUM (HTF)
Page 4 Nexus Volume 1 (June 2016) HUMAN TUBAL FLUID MEDIUM (HTF) Figure1a: SAR Healthline India (P) Ltd Figure 1b: Cryo-Genie India (P) Ltd Figure 1c: Cryocell India (P). Ltd. SINGLE DENSITY GRADIENT MEDIUM
More informationSemen Analysis Using Hamilton-Thorne Computer Assisted Semen Analyzer (CASA)
Semen Analysis Using Hamilton-Thorne Computer Assisted Semen Analyzer (CASA) 5 Ashok Agarwal, Sajal Gupta, and Rakesh Sharma 1 Introduction Semen analysis yields useful information about the fertility
More informationSystem overview Installation System Description System Default Settings and Loading I-Button Tests Components/Kits /Accessories Maintenance & Cleaning
SQA-VISION TRAINING Content of Presentation System overview Installation System Description System Default Settings and Loading I-Button Tests Components/Kits /Accessories Maintenance & Cleaning SQA-Vision
More informationThe Effect of Patient and Semen Characteristics on Live Birth Rates Following Intrauterine Insemination: A Retrospective Study 1
Journal of Assisted Reproduction and Genetics. Vol. 17, No., 000 CLINICAL ASSISTED REPRODUCTION The Effect of Patient and Semen Characteristics on Live Birth Rates Following Intrauterine Insemination:
More informationThe following protocol describes the isolation of nuclei from tissue. Item. Catalog No Manufacturer
SOP: Nuclei isolation from tissue and DNaseI treatment Date modified: 090923 Modified by: P. Sabo. (UW) The following protocol describes the isolation of nuclei from tissue. Ordering Information Item.
More informationMouse Hepatic Progenitor Organoid Culture: Supplementary Protocols
TECHNICAL BULLETIN Mouse Hepatic Progenitor Organoid Culture: The following are supplementary protocols for the culture of hepatic organoids with HepatiCult Organoid Growth Medium (Mouse) (Catalog #06030).
More informationCOMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.
This document is available at www.stemcell.com/pis EasySep Mouse Monocyte Isolation Kit Catalog #19861 For processing 1 x 10^9 cells Description Isolate untouched and highly purified monocytes from mouse
More informationTECHNICAL GUIDANCE FOR THE ACCREDITATION OF ANDROLOGY LABORATORIES
TECHNICAL GUIDANCE FOR THE ACCREDITATION OF ANDROLOGY LABORATORIES Approved By: Chief Executive Officer: Ron Josias Accreditation Executive: Mpho Phaloane Revised By: Medical Specialist Technical Committee
More informationWorksheet. Worksheet. Worksheet. Worksheet. Student Performance Guide. Student Performance Guide
LESSON 6-3 Laboratory Reagent Preparation and Calculations Worksheet LESSON 6-4 Chemistry Instrumentation in the Physician Office Laboratory Worksheet LESSON 6-6 Measuring Blood Glucose Worksheet LESSON
More informationCRYOTOP SAFETY KIT Protocol. Cryotop Method
CRYOTOP SAFETY KIT Protocol Cryotop Method R Vitrification PART Materials Required Cryotop Safety Kit-Vitrification No.0 Basic Solution (BS): 1 X 1.5ml vial (Only for Oocyte Vitrification) No.1 Equilibration
More information1.0 Purpose - This procedure specifies the method for conducting analysis for semen and sperm in forensic casework.
Procedure for Semen and Sperm Analysis 1.0 Purpose - This procedure specifies the method for conducting analysis for semen and sperm in forensic casework. 2.0 Scope - This procedure applies to those Forensic
More informationAbnormalities of Spermatogenesis
Abnormalities of Spermatogenesis Male Factor 40% of the cause for infertility Sperm is constantly produced by the germinal epithelium of the testicle Sperm generation time 73 days Sperm production is thermoregulated
More informationMagniSort Mouse CD4 T cell Enrichment Kit Catalog Number: RUO: For Research Use Only. Not for use in diagnostic procedures.
Page 1 of 2 MagniSort Mouse CD4 T cell Enrichment Kit RUO: For Research Use Only. Not for use in diagnostic procedures. Mouse splenocytes were unsorted (left) or sorted with the MagniSort Mouse CD4 T cell
More informationSQA-V Gold Semiannual Validation/Proficiency/QC Recommendations
MEDICAL ELECTRONIC SYSTEMS 5757 W. Century Blvd. Suite 805 Los Angeles, CA. 90045 Remember, it ALL Started with a Sperm! www.mes global.com SQA-V Gold Semiannual Validation/Proficiency/QC Recommendations
More informationMagniSort Human CD4 Memory T cell Enrichment Kit Catalog Number: RUO: For Research Use Only. Not for use in diagnostic procedures.
Page 1 of 2 MagniSort Human CD4 Memory T cell Enrichment Kit RUO: For Research Use Only. Not for use in diagnostic procedures. Normal human peripheral blood mononuclear cells were unsorted (left) or sorted
More informationProtn-Latex. For Determination of Protein in Latex. (Cat. # Latex, Latex) think proteins! think G-Biosciences
415PR-02 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Protn-Latex For Determination of Protein in Latex (Cat. # 786-20Latex, 786-21Latex)
More informationGlobal Histone H3 Acetylation Assay Kit
Global Histone H3 Acetylation Assay Kit Catalog Number KA0633 96 assays Version: 06 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 Principle
More informationMouse C-Peptide ELISA Kit
Mouse C-Peptide ELISA Kit Cat.No: DEIA4507 Lot. No. (See product label) Size 96T Intended Use The Mouse C-Peptide ELISA kit is for the quantitative determination of c-peptide in mouse serum, plasma, and
More informationSPERM PREPARATION, HANDLING & STORAGE
SPERM PREPARATION, HANDLING & STORAGE Solutions for the andrology laboratory HELP AT EVERY STEP with simpler processes, less stress and better results Multipurpose Handling Medium RETRIEVE AND RINSE OOCYTES
More informationMagniSort Human CD4 T cell Enrichment Kit Catalog Number: RUO: For Research Use Only. Not for use in diagnostic procedures.
Page 1 of 2 MagniSort Human CD4 T cell Enrichment Kit RUO: For Research Use Only. Not for use in diagnostic procedures. Normal human peripheral blood mononuclear cells were unsorted (left) or sorted with
More informationMagniSort Mouse CD4 Naive T cell Enrichment Kit Catalog Number: RUO: For Research Use Only. Not for use in diagnostic procedures.
Page 1 of 2 MagniSort Mouse CD4 Naive T cell Enrichment Kit RUO: For Research Use Only. Not for use in diagnostic procedures. Mouse splenocytes were unsorted (left) or sorted with the MagniSort Mouse CD4
More informationStriatal Neuron Medium Kit
Striatal Neuron Medium Kit Product Information What are included in the Striatal Neuron Medium Kit (ax0333): 2x 250 ml Striatal Neuron Basal Medium (Store at 4 o C upon receipt) 2x 7.5 ml Striatal Neuron
More informationCOMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.
This document is available at www.stemcell.com/pis Catalog #18765 EasySep Mouse CD4+CD62L+ T Cell Isolation Kit For processing 1x 10^9 cells Description Isolate highly purified naïve CD4+ T cells (CD4+CD62L+)
More informationWiCell Neural Stem Cell Protocols
WiCell Neural Stem Cell Protocols 2008 WiCell Preface This booklet of protocols is intended to serve as a primer for culturing neural stem cells in NSC culture medium. These protocols are representative
More informationChapter 30. Assessment of Oxidative Stress in Sperm and Semen. Anthony H. Kashou, Rakesh Sharma, and Ashok Agarwal. Abstract. 1.
Chapter 30 Assessment of Oxidative Stress in Sperm and Semen Anthony H. Kashou, Rakesh Sharma, and Ashok Agarwal Abstract The chemiluminescence method is the most commonly employed technique as a direct
More informationHuman Apolipoprotein A1 EIA Kit
A helping hand for your research Product Manual Human Apolipoprotein A1 EIA Kit Catalog Number: 83901 96 assays 1 Table of Content Product Description 3 Assay Principle 3 Kit Components 3 Storage 4 Reagent
More informationE.Z.N.A. SQ Blood DNA Kit II. Table of Contents
E.Z.N.A. SQ Blood DNA Kit II Table of Contents Introduction and Overview...2 Kit Contents/Storage and Stability...3 Blood Storage and DNA Yield...4 Preparing Reagents...5 100-500 μl Whole Blood Protocol...6
More information2017 SPERM QUALITY ANALYSIS MES CATALOGUE
2017 SPERM QUALITY ANALYSIS MES CATALOGUE [ 2 ] CONTENTS Page Section 4 7 Sperm quality analyzers 8 9 Digital systems for sperm analysis 15 19 Additional information [ 3 ] SPERM ANALYSIS SQA-Vision Sperm
More informationHuman TSH ELISA Kit. User Manual
Human TSH ELISA Kit User Manual Catalog number: GTX15585 GeneTex Table of Contents A. Product Description... 2 B. Kit Components... 3 C. Additional Required Materials (not included)... 3 D. Reagent Preparation...
More informationFOCUS SubCell. For the Enrichment of Subcellular Fractions. (Cat. # ) think proteins! think G-Biosciences
169PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name FOCUS SubCell For the Enrichment of Subcellular Fractions (Cat. # 786 260) think
More informationNordicCell. Andrology Phone Fax Page 1
Andrology 2013-2014 Phone +45 7025 1535 Fax +45 7025 1531 Email info@.com Page 1 www..com IVFpipette TM - improved ergonomics The IVFpipette is designed to make it possible to pipette small volumes and
More informationFERTIUP PM 1 ml / 0.5 ml - CARD MEDIUM Set
Product manual FERTIUP PM 1 ml / 0.5 ml - CARD MEDIUM Set Cat. No. KYD-004-EX Size: 1 SET KYD-005-EX 1 SET Department of Reproductive Engineering, Center for Animal Resources and Development, Kumamoto
More informationIssued by: LABORATORY MANAGER Original Date: March 20, 2000 Approved by: Laboratory Director Revision Date: July 26, 2000 CRYPTOCOCCAL ANTIGEN
Policy # MI\TECH\11\v01 Page 1 of 6 Issued by: LABORATORY MANAGER Original Date: March 20, 2000 Approved by: Laboratory Director Revision Date: July 26, 2000 CRYPTOCOCCAL ANTIGEN Latex particles coated
More informationReda Mahfouz, MD Jeffrey Goldberg MD Tommaso Falcone, MD
CENTER for REPRODUCTIVE MEDICINE Glickman Urological and Kidney Institute and Obstetrics Gynecology and Women s Health Institute Duration: August 1 30, 2008 Locations: TT3 (Cleveland Clinic), A81 Gynecology
More informationXVII Congresso Internazionale SIVE
SOCIETÀ ITALIANA VETERINARI PER EQUINI SOCIETÀ FEDERATA ANMVI XVII Congresso Internazionale SIVE XVII SIVE International Congress Palacongressi d Abruzzo Montesilvano (PE) - ITALY 4-6 Febbraio 2011 February
More informationColloid Centrifugation Selects Normal Spermatozoa from Polymorphic Bull Ejaculates: A Case Study
Colloid Centrifugation Selects Normal Spermatozoa from Polymorphic Bull Ejaculates: A Case Study J.M. Morrell, Heriberto Rodriguez-Martinez and M. Andersson Linköping University Post Print N.B.: When citing
More informationInstructions for Use. APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests
3URGXFW,QIRUPDWLRQ Sigma TACS Annexin V Apoptosis Detection Kits Instructions for Use APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests For Research Use Only. Not for use in diagnostic procedures.
More informationSemen. Dr. Mohamed Saad Daoud
Semen Dr. Mohamed Saad Daoud 1 Reference Books: Urinanalysis and body fluids (Susan King Strasinger- Marjorie Schaub De Lorenzo) Fifth edition Dr. Mohamed Saad Daoud 2 Diagram of the male genitalia Dr.
More informationItem Catalog Number Manufacturer 1,4-Dithioerythritol (1 g) D9680 Sigma-Aldrich
SOP: Nuclei isolation from fresh mouse tissues and DNaseI treatment Date modified: 01/12/2011 Modified by: E. Giste/ T. Canfield (UW) The following protocol describes the isolation of nuclei and subsequent
More informationChlamydia Rapid Screen Test (RAP-2858) RUO in the USA. Revised 28 Jul 2006
INDICATION For the rapid detection of Chlamydia Trachomatis antigens in swab specimens. For in vitro diagnostic use only, except in the United States where it is intended for Research Use Only. SUMMARY
More informationEPIGENTEK. EpiQuik Global Histone H3 Acetylation Assay Kit. Base Catalog # P-4008 PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE
EpiQuik Global Histone H3 Acetylation Assay Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik Global Histone H3 Acetylation Assay Kit is suitable for specifically measuring global
More informationHuman ipsc-derived Microglial Precursors
Human ipsc-derived Microglial Precursors Product Information Catalog. No. Product Name Format Storage on Arrival ax1666 Human ipsc-derived Microglial Precursors 3 x 10 6 viable cells Transfer cells to
More informationHuman HIV (1+2) antigen&antibody ELISA Kit
Human HIV (1+2) antigen&antibody ELISA Kit Catalog Number. CSB-E18042h For the qualitative determination of human HIV (1+2) antibody and P24 antigen concentrations in serum, plasma. This package insert
More informationRayBio Human Phospho-DDR2 (Tyr740) and Total DDR2 ELISA Kit
RayBio Human Phospho-DDR2 (Tyr740) and Total DDR2 ELISA Kit Catalog #: PEL-DDR2-Y740-T User Manual Last revised March 22, 2018 Caution: Extraordinarily useful information enclosed ISO 13485 Certified 3607
More informationRayBio Human Phosphotyrosine BTK ELISA Kit
RayBio Human Phosphotyrosine BTK ELISA Kit Catalog #: PEL-BTK-Y User Manual Last revised August 10, 2016 Caution: Extraordinarily useful information enclosed ISO 13485 Certified 3607 Parkway Lane, Suite
More informationMidi Plant Genomic DNA Purification Kit
Midi Plant Genomic DNA Purification Kit Cat #:DP022MD/ DP022MD-50 Size:10/50 reactions Store at RT For research use only 1 Description: The Midi Plant Genomic DNA Purification Kit provides a rapid, simple
More informationEPIGENTEK. EpiQuik Global Histone H4 Acetylation Assay Kit. Base Catalog # P-4009 PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE
EpiQuik Global Histone H4 Acetylation Assay Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik Global Histone H4 Acetylation Assay Kit is suitable for specifically measuring global
More informationCOMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.
This document is available at www.stemcell.com/pis Catalog #18765 EasySep Mouse CD4+CD62L+ T Cell Isolation Kit For processing 1x 10^9 cells Description Isolate highly purified naïve CD4+ T cells (CD4+CD62L+)
More informationTherapeutic Sperm Banking
Therapeutic Sperm Banking An Option for Preserving Male Fertility Andrology Laboratory and Reproductive Tissue Bank Information For Patients Men undergoing cancer treatment, including certain types of
More informationProtocol for Thawing Cryopreserved Hepatocytes
cell and tissue-based products Protocol for Thawing Cryopreserved Hepatocytes Product Instruction The following procedure may be carried out in a biosafety containment hood to reduce the risk of contamination
More informationSergey I. Moskovtsev and Clifford L. Librach
Chapter 2 Methods of Sperm Vitality Assessment Sergey I. Moskovtsev and Clifford L. Librach Abstract Sperm vitality is a re fl ection of the proportion of live, membrane-intact spermatozoa determined by
More informationRayBio Human Phospho-DDR1 (Tyr792) ELISA Kit
RayBio Human Phospho-DDR1 (Tyr792) ELISA Kit Catalog #: PEL-DDR1-Y792 User Manual Last revised March 22, 2018 Caution: Extraordinarily useful information enclosed ISO 13485 Certified 3607 Parkway Lane,
More informationTitle: Column Chromatography of Green Fluorescent Protein
Title: Column Chromatography of Green Fluorescent Protein Approvals: Preparer Date_07Oct06 Reviewer: Mary Jane Kurtz Date 09Jul13 Part I Crude Isolation of GFP from Lysed Cells q Page 1 of 6 1. Purpose:
More informationPluricyte Cardiomyocytes. using the Multiwell MEA System from Multi Channel Systems
Pluricyte Cardiomyocytes using the Multiwell MEA System from Multi Channel Systems User Guide Version 3.1 / March 2018 Contents 1. Introduction 2 2. Workflow 3 3. Important Recommendations 4 4. Equipment,
More informationCONSENSUS PROTOCOL FOR PBMC CRYOPRESERVATION AND THAWING Version 3.0 Revision dated 1/8/02 (Changes are in Red Type)
CONSENSUS PROTOCOL FOR PBMC CRYOPRESERVATION AND THAWING Version 3.0 Revision dated 1/8/02 (Changes are in Red Type) 1. EQUIPMENT/SUPPLIES/REAGENTS 1.1 Gloves (latex, vinyl, nitrile) 1.2 Lab coat or protective
More informationPluricyte Cardiomyocytes
Pluricyte Cardiomyocytes Manual Version 2.1 / March 2018 Contents 1. Introduction 2 2. Equipment, Materials and Reagents 3 3. Methods 4 3.1 Coating of tissue culture plates 4 3.2 Thawing Pluricyte Cardiomyocytes
More informationRayBio Human Phospho-DDR1 (Tyr792) and Total DDR1 ELISA Kit
RayBio Human Phospho-DDR1 (Tyr792) and Total DDR1 ELISA Kit Catalog #: PEL-DDR1-Y792-T User Manual Last revised March 22, 2018 Caution: Extraordinarily useful information enclosed ISO 13485 Certified 3607
More informationTo provide you with necessary knowledge and skills to accurately perform 3 HIV rapid tests and to determine HIV status.
Module 9 Performing HIV Rapid Tests Purpose To provide you with necessary knowledge and skills to accurately perform 3 HIV rapid tests and to determine HIV status. Pre-requisite Modules Module 3: Overview
More informationRayBio Human, Mouse and Rat Phospho-NF-kB P65 (Ser536) and Total NF-kB P65 ELISA Kit
RayBio Human, Mouse and Rat Phospho-NF-kB P65 (Ser536) and Total NF-kB P65 ELISA Kit Catalog #: PEL-NFKBP65-S536-T User Manual Last revised October 10, 2017 Caution: Extraordinarily useful information
More informationChromatin Immunoprecipitation (ChIPs) Protocol (Mirmira Lab)
Chromatin Immunoprecipitation (ChIPs) Protocol (Mirmira Lab) Updated 12/3/02 Reagents: ChIP sonication Buffer (1% Triton X-100, 0.1% Deoxycholate, 50 mm Tris 8.1, 150 mm NaCl, 5 mm EDTA): 10 ml 10 % Triton
More informationSTAT1 (ps727) (Human/Mouse) ELISA Kit
STAT1 (ps727) (Human/Mouse) ELISA Kit Catalog Number KA2171 96 assays Version: 01 Intended for research use only www.abnova.com I. INTRODUCTION STAT1 (ps727) (Human/Mouse) ELISA (Enzyme-Linked Immunosorbent
More informationCANINE CARDIAC MYOCYTE ISOLATION PROTOCOL November 2000
CANINE CARDIAC MYOCYTE ISOLATION PROTOCOL November 2000 I. Preparation for cell isolation: A. Make sure the following items are autoclaved: 1. 1-8X8 baking dish 6. 3-1 liter bottles 2. 2-500 ml beaker
More informationCryopreserved HepaRG Cells and Media Supplements
Cryopreserved HepaRG Cells and Media Supplements Catalog No. MMHPR116 Catalog No. MMADD621 Catalog No. MMADD631 Catalog No. MMADD641 Catalog No. MMADD651 Catalog No. MMADD671 FOR RESEARCH USE ONLY Not
More informationTo place an order, please visit lifelinecelltech.com or call customer service at
Human Melanocyte Cells Instruction Manual HEMn Human Epidermal Melanocytes, Neonatal HEMn-HP Human Epidermal Melanocytes, Neonatal-Highly Pigmented HEMa Human Epidermal Melanocytes, Adult HEMa-HP Human
More informationCounting Spermatozoa - Part 1 By B. W. Pickett, Ph.D. Professor Emeritus, Colorado State University
Counting Spermatozoa - Part 1 By B. W. Pickett, Ph.D. Professor Emeritus, Colorado State University A hemacytometer, originally designed to count blood cells, has been used for many years to count other
More informationSTAT3 (py705)/ Pan STAT3 (Human/Mouse/Rat) ELISA Kit
STAT3 (py705)/ Pan STAT3 (Human/Mouse/Rat) ELISA Kit Catalog Number KA2176 96 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Principle of the Assay...
More informationMitochondrial DNA Isolation Kit
Mitochondrial DNA Isolation Kit Catalog Number KA0895 50 assays Version: 01 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4 Materials
More informationHbA1c (Human) ELISA Kit
HbA1c (Human) ELISA Kit Cat. No.:DEIA3509 Pkg.Size:96T Intended use GHbA1c (Human) ELISA Kit is a sandwich enzyme immunoassay for the quantitative measurement of human GHbA1c. General Description vhemoglobin,
More informationIMMUNOFLUORESCENT STAINING FOR RESPIRATORY VIRUS DETECTION: INFLUENZA A, INFLUENZA B AND RESPIRATORY SYNCYTIAL VIRUS
IMMUNOFLUORESCENT STAINING FOR RESPIRATORY VIRUS DETECTION: INFLUENZA A, INFLUENZA B AND RESPIRATORY SYNCYTIAL VIRUS A. Introduction Influenza A, Influenza B and RSV are responsible for serious respiratory
More informationRubella Latex Agglutination Test
Rubella Latex Agglutination Test Cat. No.:DLAT1088 Pkg.Size:30T Intended use The Rubella Latex Agglutination Test is a rapid latex particle agglutination test for the qualitative and semi-quantitative
More informationHuman Thyroid-Peroxidase Antibody, TPO-Ab ELISA Kit
Human Thyroid-Peroxidase Antibody, TPO-Ab ELISA Kit Catalog No: E0442h 96 Tests Operating instruction www.eiaab.com FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH
More informationG-SERIES Manual. Recommended use of G-Series Edition 9, 2017
G-SERIES Manual Recommended use of G-Series Edition 9, 2017 2002 2017 Vitrolife Sweden AB. All rights reserved. You may copy this for internal use only, not for publishing. The Vitrolife logotype is a
More informationSerum Amyloid A ELISA
Serum Amyloid A ELISA For the quantitative determination of serum amyloid A (SAA) in serum plasma For Research use Only. Not for Use in Diagnostic Procedures Please see Appendix A for Reference Serum information
More informationARTIFICIAL INSEMINATION HOMOLOGOUS WITH OLIGOSPERMIC SEMEN SEPARATED ON ALBUMIN COLUMNS*
FERTILITY AND STERILITY Copyright 1979 The American Fertility SOciety Vol. 31, No. I, January 1979 Printed in V.SA. ARTIFICIAL INSEMINATION HOMOLOGOUS WITH OLIGOSPERMIC SEMEN SEPARATED ON ALBUMIN COLUMNS*
More informationTECHNICAL BULLETIN. Catalog Number RAB0447 Storage Temperature 20 C
Phospho-Stat3 (ptyr 705 ) and pan-stat3 ELISA Kit for detection of human, mouse, or rat phospho-stat3 (ptyr 705 ) and pan-stat3 in cell and tissue lysates Catalog Number RAB0447 Storage Temperature 20
More informationINFRAFRONTIER-I3 - Cryopreservation training course. Hosted by the Frozen Embryo and Sperm Archive, MRC - Harwell
Hosted by the Frozen Embryo and Sperm Archive, MRC - Harwell IVF recovery procedure incorporting methyl-β-cyclodextrin and reduced glutathione This protocol is based on the work published by Takeo et al.,
More informationRecipes for Media and Solution Preparation SC-ura/Glucose Agar Dishes (20mL/dish, enough for 8 clones)
Protocol: 300 ml Yeast culture preparation Equipment and Reagents needed: Autoclaved toothpicks Shaker Incubator set at 30 C Incubator set at 30 C 60 mm 2 sterile petri dishes Autoclaved glass test tubes
More informationCanine Thyroid Stimulating Hormone, TSH ELISA Kit
Canine Thyroid Stimulating Hormone, TSH ELISA Kit Catalog No: E0463c 96 Tests Operating instruction www.eiaab.com FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH
More informationAssessment of pro-arrhythmic effects using Pluricyte Cardiomyocytes. on the ACEA xcelligence RTCA CardioECR
Assessment of pro-arrhythmic effects using Pluricyte Cardiomyocytes on the ACEA xcelligence RTCA CardioECR User Guide Version 3.0 / March 2018 Contents 1. Introduction 2 2. Workflow 3 3. Important recommendations
More informationaxion Protocol Cell Culture on Microelectrode Arrays Cell Type: GE Healthcare - Cytiva TM Plus Cardiomyocytes BioSystems v. 1.0
axion BioSystems Cell Culture on Microelectrode Arrays Cell Type: GE Healthcare - Cytiva TM Plus Cardiomyocytes Protocol v. 1.0 Trademarks Axion BioSystems, Inc. and the logo are trademarks of Axion BioSystems,
More informationTECHNICAL BULLETIN. Phospho-Akt (pser 473 ) ELISA Kit for detection of human, mouse, or rat phospho-akt (pser 473 ) in cell and tissue lysates
Phospho-Akt (pser 473 ) ELISA Kit for detection of human, mouse, or rat phospho-akt (pser 473 ) in cell and tissue lysates Catalog Number RAB0011 Storage Temperature 20 C TECHNICAL BULLETIN Product Description
More informationab Exosome Isolation and Analysis Kit - Flow Cytometry, Plasma
Version 1 Last updated 25 May 2018 ab228565 Exosome Isolation and Analysis Kit - Flow Cytometry, Plasma For the isolation/detection of exosomes from human plasma, urine or cell culture media. This product
More informationApolipoprotein A-1 ELISA
Apolipoprotein A-1 ELISA For the quantitative determination of apolipoprotein A1 in serum and plasma. For Research Use Only. Not For Use In Diagnostic Procedures. Please read carefully due to Critical
More informationAmerican Society for Reproductive Medicine, Birmingham, Alabama
PRACTICE COMMITTEE FERTILITY AND STERILITY VOL. 82, NO. 6, DECEMBER 2004 Copyright 2004 American Society for Reproductive Medicine Published by Elsevier Inc. Printed on acid-free paper in U.S.A. Revised
More informationab Nuclear Extract Kit
Version 1 Last updated 10 November 2017 ab221978 Nuclear Extract Kit For the preparation of nuclear extracts from mammalian and tissue. This product is for research use only and is not intended for diagnostic
More information155 S. Milpitas Blvd., Milpitas, CA USA T: (408) F: (408)
ExoQuant TM Tumor-derived Exosome enrichment and Quantification Assay Kit (biological fluids & cell media, luminometric) (Catalog # K129-1; 1 assays (96 well transparent immunoplate for luminometric readings;
More informationPROTOCOL: OPTIMIZATION OF LENTIVIRAL TRANSDUCTION USING SPINFECTION
Last Modified: April 2018 Last Review: October 2018 PROTOCOL: OPTIMIZATION OF LENTIVIRAL TRANSDUCTION USING SPINFECTION Table of Contents 1. Brief Description 1 2. Materials and Reagents.1 3. Optimization
More informationCENTER FOR REPRODUCTIVE MEDCINE
CENTER FOR REPRODUCTIVE MEDCINE Glickman Urological and Kidney Institute and Ob/ Gyn and Women s Health Institute Duration: April 1 30, 2008 Locations: TT3 (Cleveland Clinic), A81 Gynecology ART Teaching
More informationPlasma Membrane Protein Extraction Kit
ab65400 Plasma Membrane Protein Extraction Kit Instructions for Use For the rapid and sensitive extraction and purification of Plasma Membrane proteins from cultured cells and tissue samples. This product
More informationSTAT3 (py705) (Human/Mouse/Rat) ELISA Kit
STAT3 (py705) (Human/Mouse/Rat) ELISA Kit Catalog Number KA2175 96 assays Version: 01 Intended for research use only www.abnova.com I. INTRODUCTION STAT3 (py705) (Human/Mouse/Rat) ELISA (Enzyme-Linked
More informationEXPERIMENT 13: Isolation and Characterization of Erythrocyte
EXPERIMENT 13: Isolation and Characterization of Erythrocyte Day 1: Isolation of Erythrocyte Steps 1 through 6 of the Switzer & Garrity protocol (pages 220-221) have been performed by the TA. We will be
More informationGuidelines for human andrology laboratories
Guidelines for human andrology laboratories The American Fertility Society Birmingham, Alabama I. Organization of the Laboratory and Definition of Services A. General Laboratory 1. The institutional affiliation,
More informationProcedure for Blood Analysis. 1.0 Purpose - This procedure specifies the methods for performing blood analysis in forensic casework.
Procedure for Blood Analysis 1.0 Purpose - This procedure specifies the methods for performing blood analysis in forensic casework. 2.0 Scope - This procedure applies to those Forensic Scientists who have
More informationAPOB (Human) ELISA Kit
APOB (Human) ELISA Kit Catalog Number KA4330 96 assays Version: 01 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 Principle of the Assay...
More informationVirus Harvest AAV 15 cm 2
Virus Harvest AAV 15 cm 2 1) Gently scrape cells from plate in 10 ml of media, place remaining 10 ml of media in plate lid. 2) Once cells are removed from plate, wash plate by pipetting 20 ml of media
More informationTHE CRYOTEC METHOD Manual For Oocytes, Embryos And Blastocyst Vitrification
THE CRYOTEC METHOD Manual For Oocytes, Embryos And Blastocyst Vitrification Ph: +919819855905; +91 9821618106 Email: info@cryotechjapan.com www.cryotechjapan.com VITRIFICATION PART 1 - Materials Required
More informationIn vitro human regulatory T cell expansion
- 1 - Human CD4 + CD25 + regulatory T cell isolation, Workflow in vitro expansion and analysis In vitro human regulatory T cell expansion Introduction Regulatory T (Treg) cells are a subpopulation of T
More informationXTRAKT FFPE Kit / Manual
XTRAKT FFPE Kit / Manual For manual extraction of RNA, mirna and/or DNA from Formalin Fixed Paraffin Embedded (FFPE) tissue samples Catalog No. # XTK2.0-96 For research use only. Not intended for diagnostic
More informationRODENT Hepatocytes Care Manual
RODENT Hepatocytes Care Manual INSTRUCTION MANUAL ZBM0054.02 SHIPPING CONDITIONS Rodent Hepatocytes cryopreserved Orders are delivered via Federal Express courier. All US and Canada orders are shipped
More information